Figure 2
Metabolomic investigation of PPARδ and PPARγ activation in 3T3-L1 adipocytes. (a) Chromatogram of GC-MS analysis of the total fatty acid content of 3T3-L1 adipocytes treated with the PPARδ agonist. Key metabolites are labeled. (b) Plot of partial least squares-discriminant analysis (PLS-DA) scores showing the clustering of GC-MS chromatograms from the lipid fraction of 3T3-L1 adipocytes treated with 100 nM and 1 μM PPARδ agonist GW610742 compared with the control group: 1 μM PPARδ agonist dose (diamonds; n = 6), 100 nM PPARδ agonist dose (filled circles; n = 6), control (filled squares; n = 6) (R2(X) = 77%, Q2 = 75%). (c) Plot of PLS-DA scores showing the clustering of GC-MS chromatograms from the organic fraction of 3T3-L1 adipocytes treated with 10 nM PPARγ agonist GW347845 and 100 nM PPARγ agonist GW347845 compared with the control group: 10 nM PPARγ agonist dose (asterisks; n = 6), 100 nM PPARγ agonist dose (squares; n = 6), control (filled squares; n = 6) (R2(X) = 87%, Q2 = 90%). (d) Plot of PLS-DA scores showing the clustering of DI-MS negative mode ionization chromatograms from the organic fraction of 3T3-L1 adipocytes treated with 100 nM and 1 μM PPARδ agonist GW610742 compared with the control group: 1 μM PPARδ agonist dose (diamonds; n = 6), 100 nM PPARδ agonist dose (filled circles; n = 6), control (filled squares; n = 6) (R2(X) = 70%, Q2 = 85%). (e) Plot of PLS-DA scores showing the clustering of DI-MS negative mode ionization chromatograms from the organic fraction of 3T3-L1 adipocytes treated with 10 nM PPARγ agonist GW347845 and 100 nM PPARγ agonist GW347845 compared with the control group: 10 nM PPARγ agonist dose (asterisks; n = 6), 100 nM PPARγ agonist dose (squares; n = 6), control (filled squares; n = 6) (R2(X) = 86%, Q2 = 88%). (f) Key steady state metabolic changes detected in 3T3-L1 adipocytes following treatment with the PPARδ agonist GW610742 using a combination of 1H NMR spectroscopy and GC-MS. Metabolites increased in concentration are labeled in red, and metabolites decreased in concentration are labeled in blue. (g) Key steady state metabolic changes detected in 3T3-L1 adipocytes following treatment with the PPARγ agonist GW347845 using a combination of 1H NMR spectroscopy and GC-MS. Metabolites increased in concentration are labeled in red, and metabolites decreased in concentration are labeled in blue. (h) Changes in BCAAs in the culture media of PPARδ agonist-treated 3T3-L1 cells **P < 0.005, ****P < 0.0001. Error bars represent standard errors of the mean.